Anti-inflammatory thiazole compositions and methods for using same

ABSTRACT

A broad class of thiazole derivatives, including certain 2amino-thiazoles and 2-(N-morpholido)-thiazoles, are useful as anti-inflammatory agents.

United States Patent Ariyan et al. 1 July 22, 1975 ANTl-lNFLAMMATORYTHIAZOLE [51] Int. Cl. A61K 27/00 COMPOSITIONS AND METHODS FOR [58]Field of Search 424/248 USING SAME [75] Inventors: Zaven S. Ariyan,Woodbury. Conn; {56] References Cited William A. Kulka, Guelph, CanadaUNITED TATES PATENTS 1 Assignees: y 9 Inc New York1 N'YJ, 3,547,9l712/1970 Kulka (it al, 71/90 U' lLtd.,M t LC d mmya on rea ana a PrimaryExammerStanly J. Friedman Flledi 1973 Attorney, Agent. or Firm-WillardR. Sprowls, Esq.

[21] Appl. No.1 405,944

[57] ABSTRACT Related [15. Appllcatlon Data a 0 Y, fs N 70 143 N 22 I9 PN A broad class of thlazole denvat1ves,nncludmg certain mslon 0Z-amino-thiazoles and 2-(N-morpholido)-thiaz0les US. Cl. 424/248 areuseful as anti-inflammatory agents.

28 Claims, No Drawings ANTl-INFLAMMATORY THIAZOLE COMPOSITIONS ANDMETHODS FOR USING SAME CROSS REFERENCE TO RELATED APPLICATIONS This is adivision of application Ser. No. 201,143, filed Nov. 22, 1971, now U.S.Pat. No. 3,796,800.

This application is related to, and incorporates by reference, U.S. Pat.No. 3,547,9l7 to Kulka et al. disclosing methods for preparing many ofthe thiazole derivatives of the present invention.

This application is also related to Canadian Patent 837,5l7 to vonSchmeling et al. disclosing the fungicidal and plant growth regulatingactivity of many of the thiazole derivatives of the present invention.

BACKGROUND OF THE INVENTION l. Field of the Invention The inventionrelates to thiazole derivatives which are useful as anti-inflammatoryagents, i.e., they prevent and/or inhibit the formation of granulomatissue in animals. Accordingly, the invention is, in one aspect thereof,a method of preventing and/or inhibiting the formation of granulomatissue in animal subjects. In a second aspect, the invention is a classof pharmaceutical compositions containing the present thiazolederivatives.

2. Description of the Prior Art Thiazole derivatives, including numerous2 aminothiazoles are known. However, none of the known Z-amino-thiazolecompounds has ever been disclosed as having anti-inflammatoryproperties.

British patent l,099,389 describes certain 2,4- disubstituted thiazoles(which are structurally dissimilar to the compounds of the presentinvention), e.g.,

N CI-I COOH I ll S as having anti-inflammatory activity. This compoundcontains an acidic function, and the acidic function has been postulatedas being responsible for the reactivation of latent ulcers. Thus, acidicmoieties are considered to be ulcerogenic. The compounds of the presentinvention, as will become apparent, are non-acidic compounds, and thusdo not suffer from the drawbacks of the known compounds.

The patent to Kulka et al. (3,547,917) noted above disclosed a broadclass of amino-thiazoles and methods for preparing same.

According to the Kulka et al patent, amino-thiazoles are prepared bywell known methods of thiazole synthesis. Thus, as described in Kulka etal., a thioamide of the formula:

is reacted with an a-halocarbonyl compound of the formula:

Hal- H-D in the presence of a solvent such as water or alcohol withheating, followed by basification to form a compound of the formula:

wherein A is an amino or substituted amino group, B is an alkyl group, Dis a carbamoyl or a monoor disubstituted carbamoyl group and Hal is ahalogen.

Alternatively, the thiamide may be reacted with SO Cl and anon-halogenated precursor of the rx-halocarbonyl compound, namely,

CH,D

in benzene or toluene to form the hydrochloride of the 2-aminothiazole,after which the free base is recovered therefrom.

The von Schmeling et al. patent, noted above, discloses and claimsmethods of using the said Kulka et al compounds for agriculturalpurposes.

SUMMARY OF THE INVENTION The invention provides a safe and effectivemethod of preventing and inhibiting the formation of granuloma tissue inan animal subject. This is achieved by administering to an animalsubject a therapeutically effective amount of at least one compoundselected from a very large group of thiazole derivatives, including 2-amino-thiazoles and 2-(N-morpholino)-thiazoles. Generally, the amountadministered will be from about 0.1 to I00 mg/kg/day of body weight,preferably from about 10 to 25 mg/kg/day. In humans, the amount will befrom about 0.l to 1 mg/kg/day, preferably from about 0.25 to 0.6mg/kg/day.

The invention further provides a new class of pharmaceuticalcompositions comprising said thiazole derivatives which are effective asanti-inflammatory agents.

The 2-amino-thiazoles which are among those used in the present methodsare those having the formula:

N R Rll L s R wherein R is a lower alkyl group (e.g., methyl or ethyl)and R is -H, CONHC H or CONI-IC I-I (2,6- diethyl); and pharmaceuticallyacceptable acid addition salts thereof such as the hydrochloride.

The pharmaceutical compositions according to the invention comprise, incombination, a therapeutically effective amount of at least one of theabove-described thiazole derivative and a pharmaceutically acceptablecarrier and/or diluent therefor.

For example, in the case of a tablet, the composition will comprise, inaddition to the active ingredients, fillers, binders and diluents suchas lactose, methylcellulose, talc, gum tragacanth, gum acacia, agar,polyvinylpyrrolidone, stearic acid, and/or corn starch, etc. In the caseof a liquid suspension for oral administration, the composition willcomprise, in addition to the active ingredients, a filler such as sodiumcarboxymethylcellulose and/or syrup, e.g., a glycerine based syrup. Inthe case of a parenteral solution or suspension, the composition willcomprise, in addition to the active ingredient, a suitable solvent orother liquid such as a saline solution. In the case ofa topicalointment, the composition will comprise, in addition to the activeingrediem, a vehicle such as petroleum jelly or hydrophilic petrolatum.

The most preferred compound from among all those of the formula (A) is2-amino-2',4,4',6-tetramethyl-5- thiazolecarboxanilide, i.e., thecompound wherein R is methyl, R" and R' are hydrogen, and R is CONI-IR,in which R is 2,4,6-trimethylphenyl. This compound in the rat (at a doseof 200 mg/kg) produces a reduction of 77% in carrageenin induced edema.

The most preferred compound from among all those of the formula (B) is2-(N-morpholino)-4-methyl-S- thiazolecarboxanilide, i.e., the compoundwherein R is methyl and R is CONl-IC H This compound in the rat (at adose of 200 mg/I-lg) produces a reduction of 52% in carrageenin inducededema.

DETAILED DESCRIPTION The 2-amino'thiazoles of the present invention canbe prepared by the methods disclosed in the Kulka et al. patent.

One method is the reaction sequence which comprises reacting ana-halocarbonyl compound of the formula:

N s RIII/ wherein R, R, R" and R' are as defined above and Hal is ahalogen to form the hydrohalide salt of the 2- amino-thiazole derivativeof the formula (A):

The free base is obtained from the hydrohalide salt by basification withan alkali such as ammonium hydroxide, an alkali metal hydroxide, Nal-lCOetc.

The actual preparation of the Z-amino-thiazole derivatives consists ofmixing the a-halocarbonyl compound with the thioamide (excess thioamidemay be used) in the presence of a suitable solvent such as water oralcohol and heating the mixture on the steam bath for a short time (15minutes to 2 hours) followed by basification. The precipitatedamino-thiazole is filtered off, washed with water and with benzene. Theaminothiazoles are insoluble in benzene and this enables theirpreparation from crude a-halocarbonyl compound because the impuritiespresent in such a crude starting material are usually benzene-solubleand may be washed out of the end product.

Alternatively the reaction may be carried out in one step by mixingtogether the thioamide, the unhalogenated precursor of thea-halocarbonyl compound, i.e.,

and sulfuryl chloride in benzene or toluene, heating for a short timeand then recovering the Z-amino-thiazole from its hydrohalide.

The 2-(N-morpholido)-thiazoles of the present invention can be by thesame methods as are used to prepare the 2-amino-thiazoles, except that athioamide of the formula:

is reacted with an a-halocarbonyl compound of the formula:

O C-R.

Hal-CH-R,

to form the hydrohalide salt of the 2-(N-morpholido)- thiazole of theformula (B):

wherein R R and Hal are as defined above.

As previously stated, the Kulka et al. patent discloses and exemplifiesthe preparation of many of the compounds of the present invention usingthe abovedescribed methods. For the sake of convenience and completenesshowever, there follow working examples showing the preparation of someof the present compounds using the said methods. It will be understood,of course, that the methods is also applicable to all the compounds ofthe present invention.

EXAMPLE 1 2-amino-4-methyl-5-thiazolecarboxanilide Method A 846 g ofa-chloroacetoacetanilide (4 moles), 310 g of thiourea and I400 ml ofethanol were mixed thoroughly at room temperature. An exothermicreaction soon started and solution was virtually complete within a fewminutes. The reaction flask was then placed on a steam cone and heatedfor -20 minutes. Precipitation of hydrochloride started almostimmediately. The reaction mixture was cooled and the hydrochloridecollected by filtration. The hydrochloride was dissolved in warm waterand the solution filtered and basified with ammonium hydroxide. Theprecipitated base was filtered off and dried. The crude product (815 g,87% yield) started to melt at 21 1 and finally melted with decompositionat 262. Recrystallization from ethanol gave 692 g of 2-amino-4-methyl-5-thiazolecarboxanilide which started to melt at 221 or higher. Yield:74%. (The purified base partially melts final melting point isvariable).

Method B A reaction mixture of a-chloroacetoacetanilide (528 g, 2.5mol), thiourea (I90 g, 2.5 mol) and water (1600 ml) was stirred andheated at 80-90 until the solid dissolved (about 1 hour). The hotsolution was filtered and the filtrate basified with a solution ofconcentrated ammonium hydroxide (28 to 30%) (203 ml) and water (300 ml).The white precipitate was filtered, washed with water and dried. Thewhite product melted at 220223 and weighed 526 g or 90%.

EXAMPLE 2 2-amino-2 -chloro-4-methyl-5-triazolecarboxanilide A solutionof sulfuryl chloride (33 g) in benzene ml) was added portionwise to aflask containing 0- chloroacetoacetanilide (5] g) and benzene (200 ml).After the reaction mixture had stood for 2 hours at room temperature thebenzene was removed under vacuum. To the residue, which crystallized oncooling, was added ethanol (225 ml) and thiourea (20 g). After thestrongly exothermic reaction had subsided, the mixture was heated for 10minutes on a steam cone. A slurry of the crude hydrochloride in waterwas treated with aqueous ammonia to liberate the base. The base wasrecrystallized from ethanol, in which it is only slightly soluble togive white crystals g or 86% yield) which melted at 258-259 withdecomposition.

EXAMPLES 3-l 5 These examples are directed to other Z-aminothiazolesprepared. The preparations of compounds I and 2 are shown in detail inExamples l-2 which are representative of the methods employed forpreparing the compounds set forth in the following Table I:

TABLE I 2- AMI NOTHI AZOLES l H ks I RI! l Example Cpd R R R" R' M.P. C.Yield Percent l Me CONHPl1 H H (l) 74 2 Me CONHPl1(OCl) H H 258-259d 863 3 -Me CONHPh(O-El) H H 198-200 29 4 4 Me CONHPh(2,4,6-triMe) H H243-246d 42 Me 5 5 --Me CON ph H H 176-17811 55 6 6 -Me CONHCH Ph H H143- l 45 6O 7 7 Me --CONHC H H H 238-240 55 8 8 Me CON( El) H H [59-[6245 9 9 Me --CON(l-Pl') H H 236-238 43 l0 10 -Me CON 0 H H 2l6-2l8 54 l ll l -Me CONHPh(2.6di-El) H H 206-209 l2 l2 -Me CONHPh(2.6di-Me) H H (2)l3 l3 CONHNH= CONHNH -MeCO H 300 56 l4 14 Me CONH(o-tolyl) H MeC0223-226 l5 l5 -Me CONHPh Et El ll0l I2 55 (1) Double M.P.. 222-224 and270-285 decomp. (2) Double M.P.. 249-25 l 275 decomp. In Table 1. --Me(H,; -El -C,H,; iPr -CH(CH,],; Ph C,H,.

at 222-223 and then becomes solid again. Decomposition occurs slowly asthe temperature is raised and the The following Examples l6-l9 aredirected to the preparation of 2-(N-morpho1ido)-thiazoles.

7 8 EXAMPLE 16 Table ll(onlinued Ex- Y Id4-(4-Ethyl-2-th|azolyl)morphol1ne hydrochloride ample Cpd R R MIR Q g4-Morpholinethiocarboxamide (20.0 g, 0.137 mole) 18 2.4416 90% andl-chloro-2-butanone (15.0 g, 0.141 mole) were 5 19 19 P 1 1 -1 2 72%heated in absolute ethanol (l50 ml) under reflux with r hydrochloridestirring for 2 hours. Evaporation of the solvent under in Table =-CH3;CIHaand vacuum yielded crude 4-(4-ethyl-2-thrazolyl) morpholinehydrochloride (27 84% yield). After recrystalli- The q p q of the prmvemwn have p zation from ethyl acetate-absolute ethanol, the product 0maeeutleal activity as ahtl'mflammatofy agents, effecmelted at l59l62. Asample of the hydrochlorid tive in the prevention and inhibition ofgranuloma tiswas dissolved in water and treated with excess ammo- Suefol'matioh- The activity is demonstrated by a test nium hydroxide toliberate the base, 4-(4- h 1 2- which involves the diminution ofexperimental edema thiazolyl)morpholine. which was extracted withchloroinduced the h Pe of the y the ihjeetteh Q form and isolated as apale amber oil when the solvent 5 eal'tageenln- Thls test a stamtal'dPmeedufe Whleh was evaporated. The NMR spectra of the hydrochloknown thePh3tmaeeut1eal ettride and the base were in agreement with the assignedThe Procedure Used for measunhg the mhlblttoh of Structures,carragecnin-induced edema is a modification of the Table 1 i directed to0th 2 1 method of Winter et al., Proc. Soc. Exptl. Biol. Med. "15320135prepared by i method 1 l l: 544 1962). The device used for measurementof the paw volume is an adaptation of the water displace- TAm F n mentprocedure described by Adamkiewicz et al., Can.

J. Biochem. Physiol. 33: 332 (1955). The present compounds were studiedfor their effectiveness in preventing the edema caused by theintraplantar injection of -r J r 2 (N nolphol" thlzmolcs 0.05 ml of asterile 1.0% solut|on of carrageemn. The resent compounds wereadministered orally one hour p o I N R prior to the injection of thecarrageenm Into the left 4 I hind paw of rats. At peak swelling time (3hours) the volume of edema was calculated by differential paw 5 5volumes.

We have found that many of the compounds produced significant inhibitionof induced edema in rats at E Yield a dose rate of 200 mg/kg. ample CpdPercent Table III, below describes and lists compounds of the [6 59 2formula exhibit reduction in edema in "16 l7 17 Me CONHPh l63-l65 8l%hind paw of the rat,

7. Reduction in Edema 200 'mg/kg Compounds of Formula (A) N R H i S N uvreduction P of induced No. R R" R' edema 1 Me CONHPh H H 33 2 MeCONHPh(o-Cl) H H 20 3 Me CONHPh(o-Et) H H 21 4 Me CONHPh(2.4.6-tri- H H77 Me 5 Me -con H H 23 Ph 6 -Me -CONHCH,Ph H H 3B 7 Me CONH-cyclohexyl HH 33 8 Me C0N(Er H H /CH(CH3), 9 -Me H H 38 CH(CH.1)2

10 Me con 0 H H 44 1 l Me CONHPh(2,6di-Et) H H 66 I2 Me-CONHPh(2,6di-Me) H H 64 13 CONHNH2CONHNH, MeCO H 34 I4 Me CONH(otolyllH CH;,CO 31 [5 -Mc CONHPh Et Et 29 Cpd. reduction No. R R, Edema at 200mg/kg H CONHPh CONHPh CONHPh(2.6-di-Et) hydrochloride As can be readilyseen from the foregoing Tables Ill and IV, all of the compounds of thepresent invention are effective in reducing induced edema by at least inthe rat at a dose of 200 mg/kg, except for compound l8 which is thehydrochloride of compound l7.

The compounds numbered 4, 6, 9, ll, 13 and 17 (see Tables Ill and IV)were selected for further study to determine the ED, in edema reduction.In this test, a group of normal rats was injected with carrageenin toinduce edema. Then the rats were treated with varying amounts of theabove-described six compounds, and the ED was determined.

The procedure used for measuring the inhibition of carrageenininducededema is the above-described modification of the method of Winter etal., Proc. Soc. Exptl. Biol. Med. 1 l 1: 544 1962). The device used formeasurement of the paw volume is an adaptation of the water displacementprocedure described by Adamkiewsicz et al., Can. J. Biochem. Physiol.33: 332 (195- 5). The above compounds were studied for theireffectiveness in preventing the edema caused by the intraplantarinjection of 0.05 ml of a sterile l.0% solution of carrageenin.Compounds were administered orally one hour prior to the injection ofthe carrageenin into the left hind paw of rats. At peak swelling time (3hours) the volume of edema was calculated by differential paw volumes.The ED value was obtained for each compound and is defined as that dosewhich reduced edema formation by or more compared with the mean controlresponse (parallel run) in 50% of the animals.

The results of this test are given in Table V.

TABLE V ED, vs. carrageenin Assay Compound No. Confidence Limits Dose(mg/kg) ED mg/kg 13-43) (20-92) l5-93) l8-86) (21-92) l2-30) 4 10. 30,I00, 300 6 1O, 30, I00, 300 9 I0, 30. l 1 10, 30, l3 l0, 30.

The metohd used was identical to that described above, except that theanimals used were adrenalectomized several days prior to assay. Sincethe results in the non-adrenalectomized animals were similar to thoseobtained in the adrenalectomized animals, it can be inferred that theanti-inflammatory activity of the test compounds was not caused by therelease of endogenous andrenocortical steroids.

The results of this test are summarized in Table VI:

TABLE VI ED, vs. Carrageenin Assay in Adrenalectomized Rats Compound No.Dose (mg/kg) ED mg/kg Confidence Limits 4 10, 30, I00, 300 27 (13-57) 610. 30. I00, 300 50 (23-1 l5) 9 10. 30, I00, 300 43 (20-90) ll 10,30,100, 300 78 (34-179) l7 10, 30, I00, 300 50 TABLE VII THERAPEUTICINDEX Compound LD, (mg/kg) ED Therapeutic No. 48 hrs. and 5 days (mg/kg)Index 4 800 27 35 6 800 50 18 9 400 43 l l I] 800 78 l8 I7 800 [8 40Table VIII gives the results of the test on compounds 4, 6, 9, I l andI? using varying doses to determine the ED in local vs. systemic edema.

In the development of anti-inflammatory agents it is important todistinguish between irritants, which often demonstrate anti-inflammatoryactivity by a counter irritant type of effect versus trueanti-inflammatory agents. The method selected for demonstrating the truelocal anti-inflammatory effect of the present compounds was thatdeveloped by Shanahan, R. W., Arch. int. Pharmacodyn, I: 186, I969. Thismethod utilizes the carrageenininduced paw edema and the drug isinjected locally simultaneously with the irritant substance,carrageenin, into the plantar surface of the hind paw of rats. Male ratsweighing between I00 and grams, fasted for 18 hours prior to use wereemployed in this study. The test compounds were added directly to the 1%carrageenin solution and injected in a volume of 0.5 ml into the plantartissue of the left hind paw. A group of control animals receivedcarrageenin only. Three hours later the edema was measured.Antiinflammatory or irritant effect was calculated as the percentincrease or decrease in edema between the control groups and the treatedgroups. Ten rats were used per group. The calculated ED was based on thenumber of animals in each group which showed an increase or decrease ofat least 25% change from the mean control values.

TABLE [X In the cotton pellet granuloma test, compounds 4 and 17 eachhad an ED of 3 mg/kg.

In this test. the inhibition of granuloma formation was determined by amodification of the method of Meier et al., Experientia 6:469 (I950).Essentially, the test consists of subcutaneously implanting a sterilecotton disc into rats with the concomitant oral administration of thetest compounds twice daily for four days. Removal of the pellets alongwith the granuloma formulation after five days was performed and theincrement in dry weight was considered as the measure of granulomaformation. Based on several studies, a 40% reduction in granulomaformation is considered signifcant.

Thus, a dose of 3 mg/kg is sufficient to cause a 40% reduction ingranuloma formation in 50% of the test animals.

The adjuvant-induced arthritis test was also conducted in rats usingcompound 4. This test requires one month (from to day 31). In the first17 days (Ol7), the disease is in a developing stage, while for theremainder of the month (18-31) the disease is fully de' veloped. Theresults of this test, given in terms of percent reduction of swelling inthe hind paw of the rat are shown in Table IX.

The method is essentially that of Newbould. Brit. J. Pharmacol. 21: I27,I963. The test compounds were studied in the developing arthritic stateand in the established arthritic state. Separate groups of twelve ratswere administered the compounds orally using methylcellulose as thevehicle. In the study on the developing disease. administration of thetest compounds begins on day l and on day 2 each animal is injected with.05 ml/kg of a 0.5% suspension of heat-killed Mycobacterium tuberculosisinto the plantar surface of the left hind paw. Foot volumes weremeasured by a water displacement device on the day of administration ofthe Mycobacterium and again on days 3, l0 and 17. The test compoundswere administered once daily. Body weights were recorded daily and theanimals were examined for the spread of the inflammation and the degreeof secondary lesions observed and scored as mild, moderate, or severe".For study in the established disease. another group of rats are injectedwith the Mycobacterium and foot volumes are measured and after 20 daysare again measured and administration of the test compounds begins andcontinues for l 1 days. Foot volume measurements are repeated on day 27and day 3 l. The extent of the spread of the inflammation and the degreeof lesions are recorded daily as are the body weights. The effect of thetest compounds is measured by the percentage reduction in left hind pawvolumes as compared to the hind paw volumes of the control groups.

ADJUVANT-INDUCED ARTHRITIS TEST IN RATS Reduction in Swelling Hind PawCOMPOUND 4 DAY 25 mg lg 50 mg/kg mg/kg 1, 3 43 50 59 Developing Diseasel0 I3 29 l6 I7 42 38 26 20 23 5 8 Developed Disease 27 23 I0 I4 31 25 I225 Compound 4 was further studied for its analgesic and antipyreticeffects.

Analgesic Activity Compound 4was studied in rats using the method ofRandall and Sellito (Arch. int. pharmacodyn, I l I: 406, 1957). Malerats of the Long Evans strain were used in this study. The left hind pawwas injected with 0.l ml of a 20% brewers yeast suspension. One hourlater the drug was administered orally, via stomach tube. The painthreshold was measured three hours after drug administration by applyinga steadily increasing pressure of 16 grams per second to the inflamedpaw. The end point (pain threshold) was defined as the pressurenecessary to cause the animal to struggle and/or vocalize. Compound 4was administered at the following dose levels (ten animals per group):50, 100, and 200 mg/kg. Groups of ten rats receiving the vehicle servedas controls. Phenylbutazone at a dose of 100 mg/kg was studied inparallel with compound 4. The significance of the data is based on thenumber of animals responding with a 100% increase in pain threshold. Theresults are summarized in Table X.

TABLE X The Activity of Compound 4 and Phenylbutazone in theRandall-Selina Assay Pain threshold No. of rats Inc. gms exhibiting a ofpressure 100% rise in Dose 025% Methylpain threshold Com mg/kg cellulose025% Methylcellulose pound 50 89 3/10 4 I00 98 2/10 200 I35 8/10Phenylbutazone 100 I59 10/ I 0 Inspection of the table reveals thatalthough Com pound 4 has analgesic activity it is not as active asphenylbutazone.

Antipyretic Activity and the degree of hyperthermia evaluated. Drugswere then administered and the temperatures were again taken l /ii hoursafter drug administration. The degree of antipyresis was evaluated byusing the number of animals out of five which exhibited a l fall in bodytemperature. Compound 4 at 100 and 200 mg/kg did not produce asignificant antipyresis.

The compounds of the present invention, either alone, or in the form ofpharmaceutical composition may be administered to an animal subject inany of a number of forms and via any of several routes. Thus, thecompounds or compositions thereof may be orally administered in the formof tablets, pills, capsules, or in the form of a suspension. Thecompounds may also be administered parenterally in the form of aninjectable solution or suspension. The compound or compositions thereofmay also be administered topically, in the form of an ointment orrectally, in the form of a suppository.

When orally administering the compounds or compositions, use can be madeof a tablet, pill or capsule consisting entirely of the desiredcompound, although ordinarily, a composition comprising an effectiveamount of the compound and varying amounts of one or morephysiologically inert materials such as carriers, vehicles, binders andthe like will be used. Additionally, the compounds may be orallyadministered in the form of a suspension thereof in a suitable vehiclesuch as a syrup.

When parenterally administering the compounds or compositions, use maybe made of a parenteral solution or suspension of the compound in asuitable solvent or suspension medium.

The compounds of the present invention may also be administered rectallyin the form of a suppository comprising an effective amount of thedesired compound and a suitable vehicle such as petroleum jelly.

Finally, the compounds of the present invention may be applied topicallyin the form of an ointment, salve, cream or lotion comprising aneffective amount of the desired compound and a suitable vehicle such aspetroleum jelly, etc.

The following examples are specific formulations of compositionsaccording to the invention:

EXAMPLE Tablets may be prepared by the compression of a wet granulationcontaining the following:

Dosage: l Tablet 3 times a day.

EXAMPLE 2] A liquid suspension for oral administration may be preparedin the following formulation:

Ingredients In Each 5 cc Compound No. 4 l0 mg Sodiumcarboxymethylcellulose 5 mg Syrup USP q.s. to 5 cc Dosage: l teaspoonful(5 cc) every 3 to 4 hours.

EXAMPLE 22 Dry filled capsules (DFC) consisting of two sections of hardgelatin may be prepared from the following formulation:

Ingredients In each Compound No. 4 Lactose USP q.s.

Dosage: l capsule three times a day.

EXAMPLE 23 An ointment for topical use may be prepared using thefollowing formulation:

Ingredients In each Compound No. 4 5 gm Hydrophilic petrolatum USP q.s.l00 m 8 Dosage: To be applied to inflamed skin areas as needed.

EXAMPLE 24 A parenteral suspension for intra-muscular administration maybe prepared in the following formulation:

Ingredients Compound No. 4 [0 mg isotonic solution (0.85% saline) 5 ccSurfactant (a 1% solution of polysorbate USP) l cc Having thus describedour invention, what we desire to claim and protect by Letters Patent is:

l. A method of preventing and inhibiting the formation of granulomatissue in an animal subject, said method comprising administering to ananimal subject a therapeutically effective amount of a compound hav ingthe formula as hereinafter set forth:

gig

4. A method as claimed in claim 1 wherein said compound is administeredto said animal in an orally administrable dosage form.

5. A method as claimed in claim 4 wherein said orally administrabledosage form is a pill, tablet or capsule.

6. A method as claimed in claim 4 wherein said orally administrabledosage form is a solution or suspension.

7. A method as claimed in claim 1 wherein said compound is administeredto said animal in a parenterally administrable dosage form.

8. A method as claimed in claim 7 wherein said parenterallyadministrable dosage form is a solution or sus pension.

9. A method as claimed in claim 1 wherein said compound is administeredto said animal in a topical dosage form.

10. A method as claimed in claim 9 wherein said topical dosage form isan ointment.

11. A method as claimed in claim 1 wherein said therapeuticallyeffective amount is from 0.1 to 100 mg./kg. of body weight of saidanimal per day.

12. A method as claimed in claim 11 wherein said amount is to 25 mg./kg.of body weight per day.

13. A method as claimed in claim 1 wherein said therapeuticallyeffective amount is from 0.1 to l mgJkg. of body weight of said animalper day.

14. A method as claimed in claim 13 wherein said amount is 0.25 to 0.6mg./kg. of body weight per day.

15. A pharmaceutical preparation in dosage unit form adapted foradministration to obtain an antiinflammatory effect comprising ananti-inflammatoryeffective non-toxic amount within the range from about0.1 to about 100 mg./kg. of body weight ofa compound having the formulaas hereinafter set forth:

wherein R is a lower alkyl, R is hydrogen, CONHC H or -CONHC H(2,6-diethyl); or a pharmaceutically acceptable acid addition saltthereof; in combination with a physiologically acceptable car rierand/or diluent therefor.

16. A composition as claimed in claim 15 wherein R is CH;, or C H R is Hor CONHC H or a hydrochloride thereof.

17. A composition as claimed in claim 15 wherein R is -CH;; and R5 is18. A composition as claimed in claim 15 in an orally administrabledosage form.

19. A composition as claimed in claim 18 wherein said orallyadministrable dosage form is a pill, tablet or capsule.

20. A composition as claimed in claim 19 wherein said pill, tablet orcapsule comprises 10-50 mg. of said compound.

21. A composition as claimed in claim 18 wherein said orallyadministrable dosage form is a solution or suspension.

22. A composition as claimed in claim 21 wherein said solution orsuspension comprises about 2 mg. of said compound per cc.

23. A composition as claimed in claim 15 in an injectably administrabledosage form.

24. A composition as claimed in claim 23 wherein said injectablyadministrable dosage form is a suspension.

25. A composition as claimed in claim 24 wherein said suspensioncomprises about 1.66 mg. of said compound per cc.

26. A composition as claimed in claim 15 in a topically administrabledosage form.

27. A pharmaceutical preparation in ointment form adapted foradministration to obtain an antiinflammatory effect comprising ananti-inflammatoryeffective non-toxic amount within the range from about0.1 to about l00 mg./kg. of body weight of a compound having the formulaas hereinafter set forth:

l \NVWML 0 is 5 wherein R, is a lower alkyl, R is hydrogen,

CONHC H or CONHC H (2,6-diethyl); or a pharmaceutically acceptable acidaddition salt thereof; in combination with a physiologically acceptablecarrier and/or diluent therefor.

28. A composition as claimed in claim 27 wherein said ointment comprisesabout 5 gm. of said compound per lOO gm. of total ointment.

* k IF UNITED STATES PATENT AND TRADEMARK OFFICE CERTIFICATE OFCORRECTION PATENT NO. 3,896,223 DATED July 22, 1975 WVENTOMS) I Zaven S.Ariyan; Marshall Kulka; William A. Harrison It is certified that errorappears in the aboveidentified patent and that said Letters Patent arehereby corrected as shown below:

On the cover sheet, list the inventors:

Zaven S. Ariyan, Woodbury, Conn.; Marshall Kulka, Guelph, Canada;William A. Harrison, Guelph, Canada.

Column 15, lines 35-40, the formula should appear:

Signed and Scaled this twenty-eight Day of October 1975 [SEAL] A ttes r:

RUTH C. MASON Arresting O ficer C MARSHALL DANN Commissioner nflarentsand Trademarks

1. A METHOD OF PREVENTING AN INHIBITING THE FORMATION OF GRANULOMATISSUE IN AN ANNIMAL SUBJECT, SAID METHOD COMPRISING ADMINISTERING TO ANANIMAL SUBJECT A THERAPEUTICALLY EFFECTIVE AMOUNT OF A COMPOUND HAVINGTHE FORMULA AS HEREINAFTER SET FORTH:
 2. A method as claimed in claim 1wherein R4 is -CH3 or C2H5, R5 is -H or -CONHC6H5; or a hydrochloridethereof.
 3. A method as claimed in claim 1 wherein R4 is -CH3 and R5 is-CONHC6H5.
 4. A method as claimed in claim 1 wherein said compound isadministered to said animal in an orally administrable dosage form.
 5. Amethod as claimed in claim 4 wherein said orally administrable dosageform is a pill, tablet or capsule.
 6. A method as claimed in claim 4wherein said orally administrable dosage form is a solution orsuspension.
 7. A method as claimed in claim 1 wherein said compound isadministered to said animal in a parenterally administrable dosage form.8. A method as claimed in claim 7 wherein said parenterallyadministrable dosage form is a solution or suspension.
 9. A method asclaimed in claim 1 wherein said compound is administered to said animalin a topical dosage form.
 10. A method as claimed in claim 9 whereinsaid topical dosage form is an ointment.
 11. A method as claimed inclaim 1 wherein said therapeutically effective amount is from 0.1 to 100mg./kg. of body weight of said animal per day.
 12. A method as claimedin claim 11 wherein said amount is 10 to 25 mg./kg. of body weight perday.
 13. A method as claimed in claim 1 wherein said therapeuticallyeffective amount is from 0.1 to 1 mg./kg. of body weight of said animalper day.
 14. A method as claimed in claim 13 wherein said amount is 0.25to 0.6 mg./kg. of body weight per day.
 15. A pharmaceutical preparationin dosage unit form adapted for administration to obtain ananti-inflammatory effect comprising an anti-inflammatory-effectivenon-toxic amount within the range from about 0.1 to about 100 mg./kg. ofbody weight of a compound having the formula as hereinafter set forth:16. A composition as claimed in claim 15 wherein R4 is -CH3 or -C2H5, R5is -H or -CONHC6H5; or a hydrochloride thereof.
 17. A composition asclaiMed in claim 15 wherein R4 is -CH3 and R5 is -CONCH6H5.
 18. Acomposition as claimed in claim 15 in an orally administrable dosageform.
 19. A composition as claimed in claim 18 wherein said orallyadministrable dosage form is a pill, tablet or capsule.
 20. Acomposition as claimed in claim 19 wherein said pill, tablet or capsulecomprises 10-50 mg. of said compound.
 21. A composition as claimed inclaim 18 wherein said orally administrable dosage form is a solution orsuspension.
 22. A composition as claimed in claim 21 wherein saidsolution or suspension comprises about 2 mg. of said compound per cc.23. A composition as claimed in claim 15 in an injectably administrabledosage form.
 24. A composition as claimed in claim 23 wherein saidinjectably administrable dosage form is a suspension.
 25. A compositionas claimed in claim 24 wherein said suspension comprises about 1.66 mg.of said compound per cc.
 26. A composition as claimed in claim 15 in atopically administrable dosage form.
 27. A pharmaceutical preparation inointment form adapted for administration to obtain an anti-inflammatoryeffect comprising an anti-inflammatory-effective non-toxic amount withinthe range from about 0.1 to about 100 mg./kg. of body weight of acompound having the formula as hereinafter set forth:
 28. A compositionas claimed in claim 27 wherein said ointment comprises about 5 gm. ofsaid compound per 100 gm. of total ointment.